Ectopic expression of DoFLS1 from Dendrobium officinale enhances flavonol accumulation and abiotic stress tolerance in Arabidopsis thaliana.

Abstract

Flavonols are important active ingredients that are found in abundance in Dendrobium officinale. Research on flavonol biosynthesis currently focuses on the more ubiquitous kaempferol and quercetin, but little is known on the biosynthesis of myricetin. Notably, flavonol synthase (FLS), which is responsible for the biosynthesis of flavonols, has not yet been identified. In this study, we isolated a flavonol synthase, DoFLS1, from Dendrobium officinale. DoFLS1 harbors conserved 2-oxoglutarate-dependent dioxygenase-specific and FLS-specific motifs. DoFLS1 is a cytoplasmic protein. DoFLS1 was universally expressed in roots, stems, and leaves of juvenile and adult D. officinale plants. DoFLS1 expression was strongly correlated in juvenile and adult D. officinale plants (R2 = 0.86 and 0.98, respectively; p < 0.01) with the average of corresponding flavonol levels. Transgenic Arabidopsis thaliana expressing DoFLS1 exhibited a 1.24-fold increase in flavonol content and a 25.78% decrease in anthocyanin content compare to wild-type plants, possibly resulting from a 78.61% increase in myricetin level. Moreover, the loss of anthocyanin was attributed to decreased expression of dihydroflavonol reductase (DFR) and anthocyanidin synthase (ANS) genes in transgenic A. thaliana that expressed DoFLS1. DoFLS1 also complemented the deficiency in flavonol of the A. thaliana fls1-3 mutant, which had reduced anthocyanin but increased flavonol content relative to the fls1-3 mutant. In addition, DoFLS1 was significantly upregulated after treatment with cold, drought or salicylic acid. These findings provide genetic evidence for the involvement of DoFLS1 in the biosynthesis of flavonol and in response to abiotic stresses.